Cytogenetics services include support to gene mapping, positional cloning, cytogenetic diagnostics, tumor cytogenetics, comparative evolution studies, transgenic mouse analysis, karyotyping, metaphase & interphase Fluorescent In Situ Hybridization (FISH), chromosome painting, fiber-FISH, Spectral Karyotyping (SKY), and chromosome Comparative Genomic Hybridization techniques. These services provide a spectrum of critically important techniques that are required to support the research efforts of many investigators within the institute. The culturing of cell lines, embryos, tumors, spleen and other sources of tissue are also provided. This section maintains two upright fluorescent scopes (FISH and SKY), cameras, spectracube and image analysis software. In total 1383 cytogenetic experiments were performed last year.[unreadable] Microscopy services include training investigators on how to properly use confocal laser scanning microscopy in studies which include Fluorescence Recovery After Photo-bleaching (FRAP), Fluorescent Resonant Energy Transfer (FRET), Photo-activation of Green Fluorescent Protein (PA-GFP), nuclear/organelle/cytoplasmic co-localization studies, 2-dimentional (2D), 3-dimentional (3D) & 4-dimentional (4D) cell morphology and volumetric studies, response to stimuli (drug), quantitative analysis (fluorescence, area, counts, etc), live cell and deep tissue imaging (with two-photon microscopy). The long-term live cell system gives investigators the ability to study live cells from hours to days with a temperature, humidity and CO2 controlled environment. [unreadable] This section maintains two confocal systems (UV and NLO), one long-term live-cell system, three epi-fluorescent microscopes all fitted with CCD cameras and two computer workstations. Microscopy usage for the past year; 1792 confocal hours, 4217 long-term live-cell hours, 1265 epi-fluorescent hours and 167 (only 4 months) workstation hours. [unreadable] [unreadable] The Core contributes in the following projects utilizing molecular cytogentic tools:[unreadable] Determination of chromosomal abnormalities using SKY, prior to sequence serous and clear cell endometrial tumors genomes;[unreadable] Analysis of chromosomal amplifications of specific locus in Melanoma disease progression;[unreadable] Evaluation of whole-gene deletion in Neurofibromatosis type 1 patients-cell lines and tumors touch-preps;[unreadable] Mapping of translocation breakpoints in a Hapomelanosis patient between chromosomes X and 16;[unreadable] Identification of whole-gene deletions for ATP-binding cassette, sub-family C member 6 that are the result of many segmental duplications known to lead to genomic rearrangements in Pseudoxanthoma Elasticum a rare autosomal recessive connective tissue disorder involving calcification of tissues in the retina, cardiovascular system, and flexural areas of the skin;[unreadable] Visualization of chromosome breakage in response to DNA damage caused by ELG1 protein;[unreadable] Analysis of chromosomes alterations by SKY in human disease mouse model of microcephaly;[unreadable] Studying the region of the insertion of a transgene (NSE-VEGF) in a mouse model for coloboma, a potentially blinding congenital eye malformation caused by the failure of the optic fissure to close during development;[unreadable] Screening the integration by FISH to identify putative two copies Lamin A transgenic mice;[unreadable] Screening by FISH of correct integration of modified Bacterial Artificial Chromosome DNA into the mouse embryonic stem cells for the generation of gene knockouts of different tyrosine kinases to study their role in immuno-deficiencies;[unreadable] Quantification of telomere lengths in normal and progeria cells from Hutchinson-Guilford Progeria Syndrome (HGPS) patient-cell lines;[unreadable] Comparative analysis on Indian and Chinese muntjac deer chromosomes, two species that are closely related but their genomes have drastically different organizations.[unreadable] [unreadable] The Core participates in the following projects utilizing confocal microscopy and image processing techniques:[unreadable] Immuno-flourescence studies include:[unreadable] Characterization of Neurofibromatosis type 1 (NF1); [unreadable] GFP-tagged protein foci in yeast; [unreadable] mRNA triage post-transcriptional regulation of messages in the beta cell in Type 2 Diabetes; [unreadable] Studying the relevance of a novel binding partner for the Cystic Fibrosis Trans-membrane Conductance Regulator protein (CFTR) and to elucidate its role in CFTR trafficking; [unreadable] Yes-associated protein 65 (Yap65) is a transcriptional co-activator important in cancer and development using confocal microscopy to monitor endogenous Yap translocation from the nucleus to the cytoplasm and vice-versa in response to stimuli as well as during the cell cycle in various cell types;[unreadable] Neuron-specific cyclin-dependent kinase Cdk5 is strongly implicated in the progression of many mammalian neurodegenerative diseases, including Alzheimers, Parkinson and Amyotrophic lateral sclerosis (ALS). We found that Drosophila lacking Cdk5 activator protein, p35, displays an adult-onset neurodegenerative central nervous system (CNS) syndrome. The goal of this project is to study how Cdk5/p35 causes neurodegeneration; [unreadable] The study of acute myeloid leukemia protein, Cbfb-MYH11 by confocal microscopy; examine the morphology of mitochondria from patients and mice with methylmalonic acidemia (MMA); [unreadable] Determine the sub-cellular localization of GFP-tagged TRMT12 fusion protein. Previously, we reported that the tRNA methyltransferase12 (TRMT12) gene is over-expressed in breast cancer and we are interested in studying the role of TRMT12 in tumorigenesis.[unreadable] Co-localization studies include:[unreadable] Evaluation of mechanisms in a mouse model of microcephaly; [unreadable] Signal transduction pathways utilized by the Wnt family of secreted proteins, dysregulation of this pathway has been strongly implicated in cancer and osteoporosis. The planar cell polarity pathway has been associated with spinal bifida, deafness, polycystic kidney disease and diabetes; [unreadable] How human ELG1 protein responds to multiple DNA damages and localizes where there is DNA damage using confocal microscopy and live cell imaging techniques; [unreadable] Dhx8 was identified from a zebrafish screen looking for genes involved in normal hematopoiesis and leukemia. To determine if dhx8 co-localizes with DNA or tubulin during mitosis and to observe the effect of knock-down on cell cycle progression by using confocal microscopy;[unreadable] Mutations in glucocerebrosidase gene(GBA) are seen in Gaucher disease, Parkinson disease and dementias with parkinsonism. Using several organelle markers and immunofluorescence assays for a-synuclein aggregation to show that GBA mutations lead to misfolding of protein and improper trafficking; [unreadable] Understanding the mechanisms behind the development of Autosomal Dominant Polycystic Kidney Disease by using confocal microscopy techniques to assess the role of Polycystin-2 (PC-2) binding partners on localization of PC-2 and cell-cell contacts and for co-localization with PC2.[unreadable] Fluorescent intensity measurements studies include:[unreadable] Assessing the degree of cytoplasmic sequestration of the primarily nuclear protein Runx1 through its interaction with the acute myelogenous leukemia protein, inversion 16 and its deletions mutants; [unreadable] Quantifying mucus production based on a modified mucin stain in asthma related-allergen challenged mice; [unreadable] To understand the cellular mechanisms of Hutchinson-Guilford Progeria Syndrome (HGPS) and normal aging by quantitatively measuring the telomere lengths in normal and progeria cells;[unreadable] Fluorescence Recovery After Photo-bleaching (FRAP) studies include:[unreadable] Analysis of GFP-progeria cells in mitosis; [unreadable] Quantifying the diffusion properties of the IGF2BP2 protein.